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研究生:郭雅帛
研究生(外文):Ya-Po Kuo
論文名稱:ArsA同源蛋白ARR4在酵母細胞熱耐受性表現的角色
論文名稱(外文):Role of yeast ArsA homologue ARR4 in thermotolerance of Saccharomyces cerevisiae
指導教授:許 清 玫
指導教授(外文):Ching-Mei Hsu
學位類別:碩士
校院名稱:國立中山大學
系所名稱:生物科學系研究所
學門:生命科學學門
學類:生物學類
論文種類:學術論文
論文出版年:2002
畢業學年度:90
語文別:中文
論文頁數:48
中文關鍵詞:氧化性傷害ARR4基因熱耐受性ArsA同源蛋白
外文關鍵詞:STREROSThermotoleranceARR4yeastHeat stress
相關次數:
  • 被引用被引用:2
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  • 下載下載:10
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S. cerevisiae中含有的ORF YDL100C為系統名稱,經由SGD (Saccharomyces Genome Database) 資料顯示,目前此基因名稱暫定為ARR4基因,與E. coli的ArsA基因序列經由Genetic Computer Group (GCG)序列比對,其同源性約29 %。其基因序列分析產物 ARR4 蛋白上也具有相似的ATP結合位,此結合位在胺基酸序列的第25到32的氨基酸上。之前研究指出此ATP結合位並不受陰離子激發,所以被認為和砷化物抗性機制無直接關係或無關,而此點目前也有報告指出酵母菌對砷化物的抗性另有其他機制。
本論文探討ARR4基因在酵母菌於高溫時調節細胞存活的機能上扮演的角色,先前研究初步發現ARR4基因缺失的酵母細胞在高溫下無法生長。本論文的研究結果顯示,細胞中氧化自由基的堆積過多及抗氧化能力的下降,是此ARR4基因缺失的酵母細胞在高溫下細胞致死的原因,但細胞於30℃則無此現象。由於heat stress 、oxidative stress及osmatic stress等外壓發生時會啟動STRE (stress response element ) 調控機制使細胞度過危機。進一步以RT-PCR 分析CTT1、SOD1、TSL1基因的表現與ARR4基因缺失的相關性時,結果顯示,ARR4基因缺失的突變株其CTT1基因在heat stress 24小時後mRNA的表現量比野生株有減少的現象,說明ARR4基因蛋白在酵母細胞在遇到外來壓力時可能並參與活化STRE的上游作用角色。
The ArsA homologue ARR4 in Saccharomyces cerevisiae, encoded by YDL100C. Homologues of the E. coli ArsA are found in S. cerevisiae about 29 % from Genetic Computer Group (GCG). The ARR4 gene product contains an ATP binding site that is similar to protein ArsA from E. coli
Disruption of ARR4 in yeast is not lethal but the disrupted strain was unable to grow at 40℃, suggesting that the possible cause of cell death in KO strain at 40℃ was investigated. The accumulation of trehalose and the in vivo molecular oxidation level are higher in KO strain than that in WT strain under heat stress condition. These suggest that the increased reactive oxygen species (ROS) but not the amount of thermoprotectant trehalose is most likely to be the reason for cell death in KO strain. In this report ROS scavenger system show that the activities of ROS scavenger system are lower in KO compared to that in WT strain at 30℃ or 40℃. This suggests that ARR4 is involved in the heat stress 、oxidative stress and osmatic stress triggers activation of the STRE ( stress tolerance response element) regulon.
Further studies involvement ARR4 of CTT1, SOD1, and TSL1 gene of STRE-drive gene by RT-PCR. Here the report that the KO strain exhibits a thermosensitivity phenotype in comparison to wild-type strain, indicating that ARR4 may act as a component of a stress tolerance network.
一、中文摘要……………………………………… 3
二、英文摘要……………………………………… 4
三、背景介紹……………………………………… 5
四、材料及實驗方法……………………………… 11
五、實驗結果……………………………………… 17
六、討論…………………………………………… 20
七、圖表…………………………………………… 24
八、參考文獻……………………………………… 46
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